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. 2014 May 7:12:10.
doi: 10.1186/1477-9560-12-10. eCollection 2014.

Thrombin based gelatin matrix and fibrin sealant mediated clot formation in the presence of clopidogrel

Joseph F Dwyer  1 , Jill A McCoy  1 , Ziping Yang  1 , Michael Husser  1 , Heinz Redl  2 , Mary Ann Murphy  1 , Martin Wolfsegger  3 , James P DiOrio  1 , Andreas Goppelt  3 , Shane Donovan  1
Affiliations

Thrombin based gelatin matrix and fibrin sealant mediated clot formation in the presence of clopidogrel

Joseph F Dwyer et al. Thromb J. .

Abstract

Background: Platelet inhibitors are commonly used to reduce the risk of atherothrombotic events. The aim of this study was to determine the impact of platelet inhibitors, specifically clopidogrel and aspirin, on clot kinetics, strength, and/or structure during the use of thrombin based gelatin matrices and fibrin sealants.

Methods: Blood was collected and heparinized from donors on clopidogrel (and aspirin) and age matched control donors. Blood component analysis, whole blood platelet aggregometry, and activated clotting time (ACT) were used to monitor compliance to therapy and identify any differences between donor groups. Clot kinetics and strength were analyzed using thrombelastography (TEG). Field Emission Scanning Electron Microscopy (FESEM) was used to analyze clot structure.

Results: Blood component profiles were similar for both donor groups. Aggregometry indicated that aggregation response to adenosine diphosphate (ADP) for clopidogrel donors was 12% of that for the controls (p = 0.0021), an expected result of clopidogrel induced platelet inhibition. However, blood from both donor groups had an elevated thrombin induced aggregation response. Heparinization of donor blood resulted in similarly elevated ACTs for both donor groups. TEG results indicated similar clot kinetics and strength between clopidogrel and control donor groups for blood alone and when clotting was induced using thrombin based gelatin matrices and fibrin sealants. FESEM images supported TEG findings in that similar morphologies were observed in ex vivo formed clots from both donor groups when thrombin based gelatin matrices and fibrin sealants were used.

Conclusion: These results suggest that platelet inhibitors do not negatively impact clot kinetics, strength, and structure when clotting is initiated with thrombin based gelatin matrices and fibrin sealants.

Keywords: Clopidogrel; Floseal; Hemostasis; Thrombelastography; Thrombin; Tisseel.

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Figures

Figure 1
Figure 1
Blood component analyses results. The left panel shows dot-plots of all blood parameters and group. The right panel shows a CI plot where blood parameters are indicated on the y-axis and the ratios of averages (clopidogrel relative to control) and corresponding two-sided 95% CIs on the x-axis.
Figure 2
Figure 2
Whole blood aggregation results after activation with ADP or thrombin. The left panel shows dot-plots of whole blood platelet aggregation impedance per group for aggregation induced with ADP and thrombin. The right panel shows a CI plot where the type of induction is indicated on the y-axis and the ratios of averages (clopidogrel relative to control) and corresponding two-sided 95% CIs are presented on the x-axis. No bounded two-sided 95% CI for the ratio regarding ADP is available as the numerator of the ratio (i.e. mean of clopidrogel) is not statistically different from zero.
Figure 3
Figure 3
ACT results pre and post-heparinization. The left panel shows dot-plots of activated clotting time per group and pre and post-heparinization. The right panel shows a CI plot for pre- and post-heparinization (y-axis) where the ratios of averages (clopidogrel relative to control) and corresponding two-sided 95% CIs are presented on the x-axis.
Figure 4
Figure 4
TEG data: citrated blood. The left panel shows dot-plots of all TEG parameters per donor and group. The right panel shows a CI plot where TEG parameters are indicated on the y-axis and the ratio of averages (clopidogrel relative to control) and corresponding two-sided 95% CIs on the x-axis.
Figure 5
Figure 5
TEG data: thrombin based gelatin matrix. The left panel shows dot-plots of all TEG parameters per donor and group. The right panel shows a CI plot where TEG parameters are indicated on the y-axis and the ratio of averages (clopidogrel relative to control) and corresponding two-sided 95% CIs on the x-axis. Due to the rapid rate of clot formation when using the thrombin based gelatin matrix, the TEG is not sensitive enough to generate values for R and K other than the lowest possible data value for most sample replicates preventing comparison these parameters between donor groups.
Figure 6
Figure 6
TEG data: fibrin sealant. The left panel shows dot-plots of all TEG parameters per donor and group. The right panel shows a CI plot where TEG parameters are indicated on the y-axis and the ratio of averages (clopidogrel relative to control) and corresponding two-sided 95% CIs on the x-axis. Due to the rapid rate of clot formation when using the fibrin sealant, the TEG is not sensitive enough to generate values for R other than the lowest possible data value for most sample replicates preventing comparison this parameters between donor groups.
Figure 7
Figure 7
Representative images of thrombin based gelatin matrix clots acquired with FESEM. Control (A,B), clopidogrel (C,D); 10 μm scale bars (A,C), 5 μm scale bars (B,D); Arrows: Gelatin (a), Fibrin (b), Red Blood Cells (c). No qualitative differences were observed in the fibrin structure (porosity, fiber thickness, and branching) or the cellular accumulation around the gelatin granules including large numbers of trapped RBCs between clopidogrel and control donor groups.
Figure 8
Figure 8
Representative images of fibrin sealant clots acquired with FESEM. Control (A), clopidogrel (B); 5 μm scale bars; Arrows: Fibrin (a), Red Blood Cells (b). As seen in the images of the thrombin based gelatin matrix clots, no qualitative differences were observed between clopidogrel and control donor groups. The density of the fibrin structure seen compared to the gelatin based hemostat clots may be due to the additional fibrinogen supplied by the fibrin sealant.
See this image and copyright information in PMC

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